Fatty Acıd and Essentıal Oıl Composıtıon of Echınacea Purpurea (L.) Moench, Growıng in Kahramanmaras-Turkey
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International Conference on Environmental and Biological Sciences (ICEBS'2012) December 21-22, 2012 Bangkok (Thailand) Fatty Acıd and Essentıal Oıl Composıtıon of Echınacea Purpurea (L.) Moench, Growıng in Kahramanmaras-Turkey Emel Diraz1,Sengul Karaman 2, Nilufer Koca 3 proposed mechanism of action relating to its reported ability to Abstract—Echinacea is a hardy perennial plant indigenous to stimulate to immune system [5].The immuno-stimulating effect North America, which belongs to Asteraceae or Compositae, have is caused by four reactions; activiating phagocytoses, medical properties. Echinacea purpurea (L). Moench cultivated in stimulating the fibroblast, increasing respiratory activity and Avşar Campus-Kahramanmaraş, gathered in Agust 2008. Seeds of increasing mobility of leucocytes [6]. According to the present the plants extracted for fatty acid composition with two different solvent (hexan and petroleumether) by using soxhalet apparatus, knowledge, genus Echinacea has identified seven groups of essential oils of the flowers were extracted by hydrodistillation. Fatty medically important components including polysaccaharides, acid and essential oil compositions were determined with GC-MS. flavonoids, caffeic acid derivates, essential oils, Principal fatty acid components with petroleumether and hexan polyacetylenes, alkylamides and miscellaneous chemicals [2] respectively, oleic acid % 48, % 29, palmitic acid % 16.6, % 9.2, which have been analysed applying various chromatographic linoleic acid % 13.3, % 51, stearic acid % 5, % 2.5. For essential oils germacrene-D % 11.3, caryophylleneoxide % 8.7, β-caryophyllene % separation techniques [7]. 7.2 α-cadinol % 6.3, naphthalene % 3.3, α-phellandrene % 2.9 were found main compounds. II. MATERIALS AND METHODS Essantiol oil and seed oil analyses determined with GC-MS. Keywords—Echinacea purpurea, fatty acid, essential oil, GC-MS analysis were conducted in the Plant Physiology GC-MS Laboratory in Biology Dept. of Kahramanmaras Sutcu Imam University. Qualification of the oil was analyzed on Agilent I. INTRODUCTION 5975C Mass Spectrometer coupled with Agilent GC-6890II series. CHİNACEA is a hardy perennial plant indigenous to E North America, which belongs to Asteraceae or A. Essential Oil Analysis Compositae plant family and includes nine different species Dried plant material (20g Echinacea purpurea L. Mornch ) (Echinacea purpurea, Echinacea angustifolia, Echinacea was subjected to hydro-distillation method with 4h using a pallida, Echinacea atrorubens, Echinacea laevigata, Clevenger type apparatus for determining the oil content. The Echinacea paradoxa, Echinacea songuinea, Echinacea oil compositions were determined with GC-MS. The GC was simulata, Echinacea tenensseensis) [1]. Three species of equipped with HP-88 capillary column (100 m x 250 µm x Echinacea (Echinacea purpurea, Echinacea angustifolia, 0.20 µm film thickness) and He was used carrier gas with flow Echinacea pallida) have medical properties [2] and are rate of 1.0 mL/min. The GC oven temperature was o o o commercially traded as medicinal plants and used for programmed as follows: 70 C (1 min), 230 C at of 10 C/min and then kept at 230 oC at 20 min. The injector temperature floriculture. o Currently, extracts and whole plant products made from was 250 C. The mass spectrometer was operating in EI mode Echinacea purpurea, Echinacea angustifolia, Echinacea at 70 eV. Split ratio was 20:1. Mass range 35-400m/z; scan µ pallida comprise one of the largest sectors of the several speed (amu/s): 1000. 10 L of the oil was mixed 0.5 ml µ billion dolar herbal medicine market, in North America as well dichlora methan and 1 L of the concentrations injected into the column. The components of the oil were identified by as Europe. Annual sales of Echinacea products have been comparing their retention indices and mass spectra with those estimated $ 300 milion in the US alene [3]. İncreasing market of pure authentic samples and NIST98, Willey7n.1 and demand for Echinacea products has led to a rapid expantion of Flavor2 libraries reference compounds. Echinacea cultivation [4].Echinacea is used most widely as prevention or treatment for the commen cold, with the B. Seed Oil Analysis Echinacea purpurea L. Mornch seeds 15 g were extracted Emel1. Dıraz1 Kahramanmaraş Sütçüimam University, Kahramanmaras, 46100, Turkey (phone: 00903442801400 ; fax: with petroleum ether (150 ml) and the other 15 g seeds were 00903442191042 ; e-mail: [email protected] . extracted with hexan (150 ml) for 6h using a soxhlet apparatus. The oil compositions were determined with GC-MS. The GC 35 International Conference on Environmental and Biological Sciences (ICEBS'2012) December 21-22, 2012 Bangkok (Thailand) was equipped with HP-88 capillary column (100 m x 250 µm x components with 57 % percentage. In the study of Agarwal et 0.20 µm film thickness) and He was used carrier gas with flow al. [10] the most abundant terpene found in E. Purpurea oil rate of 1.0 mL/min. The GC oven temperature was was germacrene-D, which showed a rise from 7.2 %- 33.5 % programmed as follows: 170 (1 min), 220 oC at of 10 oC/min June to December. In Hudaib et al. reseach [11] germacrene-D and then kept at 230 oC at 15 min. The injector temperature was found 57.8 % ratio. In the report of Vaverkova et al. [12] was 250 oC. The mass spectrometer was operating in EI mode germacrene-D comprises the 4.8 % of the total oil. In our at 70 eV. Split ratio was 20:1. Mass range 35-400m/z; scan study, germacrene-D (11.3 %) has the biggest peak area. Kan speed (amu/s): 1000. Echinacea purpurea L. Mornch seeds et al. [13] analysed two different Echinaceae species oil was (0.1g) vortexted with hexane (5ml) 5 min, then (Echinacea purpurea and Echinacea pallid) in Turkey under vortexed with 2N KOH (0.5ml) 5 min.and centrifuge 5 min. the organic farmer conditions and determined the essential oils 1μl of the concentrations injected into the column. The of these species and the major components in E. components of the oil were identified by comparing their purpurea were germacrene D (11.20%), spathulenol (8.83%), retention indices and mass spectra with those of pure authentic caryophyllene oxide (8.46%) and 1,5-epoxysalvial-4(14)-ene samples and wiley7n.1, Famdbwax.L, Famedb23.L, libraries (5.76%). Kan et al. and our findings are similar. reference compounds. TABLE I. III.RESULTS ESSENTIAL OILS OF FLOWERHEADS peak Retention Component Area % time 1 11.662 β-pinene 1.3 2 12.002 α-phellandrene 2.9 3 13.357 p-cymene 2.6 4 16.046 β-elemene 2.1 5 16.674 β-caryophyllene 7.2 6 17.049 α-cadinene 1.0 7 17.43 Naphthalene, 1,2,3,5,6,8a-hexahydro-4,7- 3.3 dimethyl-1-(1-ethylethyl)-,(1S-cis)- 8 17.729 Germacrene-D 11.3 9 19.733 α-farnasene 1.0 10 20.112 1,5 epoxysalvia-4(14)ene 3.3 11 21.44 Naphthalene, 1,2,3,4,4a,7-hexahydro-1,6- 1.6 Fig.1. Major Components Of Flowerheads Essential Oils dimethyl-4-(1-methylethyl)- 12 21.613 α-bisabolene 2.3 13 22.138 Bicyclo[4.4.0]dec-1-ene, 2-isopropyl-5- 1.8 The essential oil and fatty acids contents of Echinacea methyl-9methylene- purpurea were analyzed. The analyse of essential oils from the 14 22.346 Caryophyllene oxide 8.7 flowerheads was given in table I. The essential oil content of 15 22.613 γ- cadinene 2.0 E. purpurea flowerheads was 1.5 % (w/v). The essential oil 16 22.962 α-cadinol 6.3 was separeted to 89 components, 63 of them were identified, 17 23.267 Decatone 1.5 comprising 92.8% of the total yield (Table I). 18 23.434 Ethyl oleate 1.6 19 23.653 Vulgarol. B 1.9 The major components of the E. Purpurea are germacrene- 20 23.916 Aromadendrene oxide 1.1 D (11.3%), caryophyllene oxide (8.7 %), β caryophyllene (7.2 21 24.596 3,4-difloro-4 methoxybiphenyl 2.7 %), α-cadinol (6.3 %). The essential oil content was 22 24.867 Isoaromadendieneepoxide 1.9 monoterpene hydrocarbons, oxygenated monoterpens, 23 25.306 Trans (Z)- α-bisabolene epoxide 2.3 sesquiterpens and oxtgenated sesquiterpens. 5.4 % of the E. 24 26.072 Benzenepropanoic acid, octadecyl ester 1.2 25 26.104 Diepi-.alpha.-cedrene epoxide 1.2 Purpurea oil belongs to monoterpene hydrocarbons such as α- Those lower than 1% were excluded. pinene, camphane, β-pinene, myrcene, limonene, p-cymene, thujone. β-caryophyllene, α-cadinene, germacrene-D, α- farnesene, α-bisabolene, isospathulenol, were sesquiterpene hydrocarbons comprised 22.8 %, caryophyllene oxide, aromadendrene oxide and isospathulenol were oxygenated sesquiterpens comprised 10.4 % of the total oil content. Myrtenol was an oxygenated monoterpene found % 0.2 ratio. In earlier research by Mazi and Cottrell [8] E. Purpurea, E. Angustifolia and E. Pallida roots, stems, leaves and flowers oils were analyzed. Terponoids of head space of flowers and stems were about 81 %- 91%. In the study by Salehi et al.[9] oxygenated monoterpene was not found in the E. Purpurea oil and 6.4 % monoterpene hydrocarbons, 70.9 % sesquiterpene Fig. II Most Abundant Fatty Acids In Seeds With Different Solvents hydrocarbons and 15.4 % oxygenated sesquiterpens were found. Germacrene-D was the highest ratio among the 36 International Conference on Environmental and Biological Sciences (ICEBS'2012) December 21-22, 2012 Bangkok (Thailand) TABLE II. of the plant. FATTY ACIDS OF SEEDS Echinacea has shown good adaptation to Mediterranian region. pe minute component Area % ak Petroleum n-hexane REFERENCES ether 1 10.766 Oxocol-C13 0.1 - [1] R.L., McGregor, “The taxonomy of the genus Echinacea (Compositae)”, 2 10.911 Myristic acid-C14:0 0.3 0.2 The University of Kansas Sci.